The apparent specificity of NotI (5'-GCGGCCGC-3') is enhanced by M.FnuDII or M.BepI methyltransferases (5'-mCGCG-3'): cutting bacterial chromosomes into a few large pieces

Gene
B Q QiangM Nelson

Abstract

The restriction endonuclease (ENase) NotI is blocked by methylation within its recognition sequence at 5'-GCGGCmCGC-3'. This sensitivity to methylation can be used to enhance the specificity of NotI in vivo and in vitro. Modification by M.FnuDII or M.BepI methyltransferases (MTase) (5'-mCGCG-3') will block NotI (5'-GCGGCCGC-3') cleavage at overlapping MTase/ENase sites 5'-CGCGGCCGC-3' (equivalent to 5'-GCGGCCGCG-3'), and increase the apparent cleavage specificity of NotI about twofold. This 'cross-protection' procedure reduces the number of NotI fragments in the genomes of Escherichia coli and Bacillus subtilis, as resolved by pulsed field electrophoresis. Application of this method to large DNAs in vitro requires the preparation of highly purified DNA MTases.

References

Jan 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·M D Weil, M McClelland
Aug 11, 1987·Nucleic Acids Research·M McClellandM Nelson
Jan 1, 1987·Methods in Enzymology·M Nelson, M McClelland
Sep 1, 1988·Gene Analysis Techniques·J Hanish, M McClelland
Dec 25, 1988·Gene·K D LunnenD O Nwankwo
Jun 12, 1987·Science·C L SmithC R Cantor
Apr 11, 1980·Nucleic Acids Research·A P Bird
Dec 11, 1982·Nucleic Acids Research·M McClelland, R Ivarie

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Citations

Aug 11, 1991·Nucleic Acids Research·H ShuklaS M Weissman
Feb 15, 2005·Analytical Biochemistry·Yaroslav Buryanov, Taras Shevchuk
Sep 3, 2013·Journal of Hepatology·Reiner WiestMarkus Geuking
Jun 1, 1994·FEMS Microbiology Reviews·S T Cole, I Saint Girons

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